Docosahexaenoic Acid Modulates Paracellular Absorption of Testosterone and Claudin-1 Expression in a Tissue-Engineered Skin Model

Healthy skin moLEdels produced by tissue-engineering often present a suboptimal skin barrier function as compared with normal human skin. Moreover, skin substitutes reconstructed according to the self-assembly method were found to be deficient in polyunsaturated fatty acids (PUFAs). Therefore, in this study, we investigated the effects of a supplementation of the culture media with docosahexaenoic acid (DHA) on the barrier function of skin substitutes. To this end, 10 μM DHA-supplemented skin substitutes were produced (n = 3), analyzed, and compared with controls (substitutes without supplementation). A Franz cell diffusion system, followed by ultra-performance liquid chromatography, was used to perform a skin permeability to testosterone assay. We then used gas chromatography to quantify the PUFAs found in the epidermal phospholipid fraction of the skin substitutes, which showed successful DHA incorporation. The permeability to testosterone was decreased following DHA supplementation and the lipid profile was improved. Differences in the expression of the tight junction (TJ) proteins claudin-1, claudin-4, occludin, and TJ protein-1 were observed, principally a significant increase in claudin-1 expression, which was furthermore confirmed by Western blot analyses. In conclusion, these results confirm that the DHA supplementation of cell culture media modulates different aspects of skin barrier function in vitro and reflects the importance of n-3 PUFAs regarding the lipid metabolism in keratinocytes.

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Modulation of the Lipid Profile of Reconstructed Skin Substitutes after Essential Fatty Acid Supplementation Affects Testosterone Permeability

Cells ◽

10.3390/cells8101142 ◽

2019 ◽

Vol 8 (10) ◽

pp. 1142 ◽

Cited By ~ 4

Author(s):

Mélissa Simard ◽

Pierre Julien ◽

Julie Fradette ◽

Roxane Pouliot

Keyword(s):

Benzoic Acid ◽

Human Skin ◽

Barrier Function ◽

Percutaneous Absorption ◽

Skin Barrier ◽

Skin Barrier Function ◽

Skin Permeability ◽

Skin Substitutes ◽

Reconstructed Human Skin

Skin models with efficient skin barrier function are required for percutaneous absorption studies. The contribution of media supplementation with n-3 and n-6 polyunsaturated fatty acids (PUFAs) to the development of the skin barrier function of in vitro skin models remains incompletely understood. To investigate whether PUFAs, alpha-linolenic acid (ALA, n-3 PUFA) and linoleic acid (LA, n-6 PUFA), could enhance the impermeability of a three-dimensional reconstructed human skin model, skin substitutes were produced according to the self-assembly method using culture media supplemented with either 10 μM ALA or 10 μM LA. The impact of PUFAs on skin permeability was studied by using a Franz cell diffusion system to assess the percutaneous absorption of testosterone and benzoic acid. Our findings showed that ALA supplementation induced a decrease in the absorption of testosterone, while LA supplementation did not significantly influence the penetration of testosterone and benzoic acid under present experimental conditions. Both ALA and LA were incorporated into phospholipids of the skin substitutes, resulting in an increase in n-3 total PUFAs or n-6 total PUFAs. Collectively, these results revealed the under-estimated impact of n-3 PUFA supplementation as well as the importance of the n-6 to n-3 ratio on the formation of the skin barrier of in vitro reconstructed human skin models.

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Characterization of Epidermal Lipoxygenase Expression in Normal Human Skin and Tissue-Engineered Skin Substitutes

Journal of Histochemistry & Cytochemistry ◽

10.1369/0022155418788117 ◽

2018 ◽

Vol 66 (11) ◽

pp. 813-824 ◽

Cited By ~ 1

Author(s):

Carolyne Simard-Bisson ◽

Lorraine Andrée Parent ◽

Véronique J. Moulin ◽

Bernard Fruteau de Laclos

Keyword(s):

Human Skin ◽

Expression Profiles ◽

Skin Barrier ◽

Epidermal Differentiation ◽

Skin Barrier Function ◽

Skin Substitutes ◽

Differentiation Markers ◽

Normal Human Skin ◽

Normal Human ◽

Engineered Skin Substitutes

Lipoxygenases (LOXs) are enzymes likely to be involved in corneocyte lipid envelope formation and skin barrier function. In humans, mutations in epidermis-type lipoxygenase 3 ( eLOX-3) and 12R–lipoxygenase ( 12R-LOX) genes are associated with autosomal recessive congenital ichthyosis (ARCI), whereas deletion of these genes in mice causes epidermal defects. LOXs also represent a matter of interest in psoriasis as well as in cancer research. However, their expression as well as the exact role of these enzymes in normal human skin have not been fully described. Our goal was to characterize the expression of epidermal LOXs in both normal human skin and Tissue-Engineered Skin Substitutes (TESS) and to consider TESS as a potential model for LOX functional studies. Staining for epidermal differentiation markers and LOXs was performed, in parallel, on normal human skin and TESS. Our results showed similar expression profiles in TESS when compared with native skin for e-LOX3, 12R-LOX, 12S-lipoxygenase (12S-LOX), and 15-lipoxygenase 2 (15-LOX-2) but not for 15-lipoxygenase 1 (15-LOX-1). Because of their appropriate epidermal differentiation and LOX expression, TESS represent an alternative model for future studies on LOX function.

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Transepidermal Water Loss Does Not Correlate with Skin Barrier Function In Vitro

Journal of Investigative Dermatology ◽

10.1046/j.1523-1747.2002.01760.x ◽

2002 ◽

Vol 118 (5) ◽

pp. 871-875 ◽

Cited By ~ 74

Author(s):

Robert P. Chilcott ◽

Christopher H. Dalton ◽

Andrew J. Emmanuel ◽

Ceri E. Allen ◽

Simon T. Bradley

Keyword(s):

Water Loss ◽

Barrier Function ◽

Skin Barrier ◽

Transepidermal Water Loss ◽

Skin Barrier Function

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Effect of Physical and Chemical Hair Removal Methods on Skin Barrier Function in vitro: Consequences for a Hydrophilic Model Permeant

Skin Pharmacology and Physiology ◽

10.1159/000493168 ◽

2018 ◽

Vol 32 (1) ◽

pp. 8-21 ◽

Cited By ~ 5

Author(s):

Astrid Pany ◽

Victoria Klang ◽

Marion Brunner ◽

Johanna Ruthofer ◽

Elisabeth Schwarz ◽

...

Keyword(s):

Barrier Function ◽

Skin Barrier ◽

Hair Removal ◽

Skin Barrier Function ◽

Physical And Chemical

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718 Impact of essential fatty acids on the skin barrier function of tissue engineered skin substitutes

Journal of Investigative Dermatology ◽

10.1016/j.jid.2018.03.727 ◽

2018 ◽

Vol 138 (5) ◽

pp. S122

Author(s):

M. Simard ◽

R. Pouliot

Keyword(s):

Fatty Acids ◽

Barrier Function ◽

Essential Fatty Acids ◽

Skin Barrier ◽

Skin Barrier Function ◽

Skin Substitutes ◽

Engineered Skin Substitutes

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Insights into skin barrier function and its perturbation with the use of in vitro stratum corneum lipid monolayers

Journal of the American Academy of Dermatology ◽

10.1016/j.jaad.2011.11.165 ◽

2012 ◽

Vol 66 (4) ◽

pp. AB37

Keyword(s):

Stratum Corneum ◽

Barrier Function ◽

Skin Barrier ◽

Skin Barrier Function ◽

Lipid Monolayers ◽

Stratum Corneum Lipid

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Partially Hydrolysed Whey-Based Infant Formula Improves Skin Barrier Function

Nutrients ◽

10.3390/nu13093113 ◽

2021 ◽

Vol 13 (9) ◽

pp. 3113

Author(s):

Sébastien Holvoet ◽

Sophie Nutten ◽

Lénaïck Dupuis ◽

Dominique Donnicola ◽

Tristan Bourdeau ◽

...

Keyword(s):

Barrier Function ◽

Protein Hydrolysate ◽

Skin Barrier ◽

Transepidermal Water Loss ◽

Skin Barrier Function ◽

Primary Keratinocytes ◽

Total Ige ◽

Expression Of Genes ◽

Human Primary Keratinocytes

Specific partially hydrolysed whey-based infant formulas (pHF-W) have been shown to decrease the risk of atopic dermatitis (AD) in infants. Historically, AD has been associated primarily with milk allergy; however, defective skin barrier function can be a primary cause of AD. We aimed to ascertain whether oral supplementation with pHF-W can improve skin barrier function. The effect of pHF-W was assessed on transepidermal water loss (TEWL) and antibody productions in mice epicutaneously exposed to Aspergillus fumigatus. Human primary keratinocytes were stimulated in vitro, and the expression of genes related to skin barrier function was measured. Supplementation with pHF-W in neonatal mice led to a significant decrease in TEWL and total IgE, but not in allergen-specific antibody levels. The whey hydrolysate was sufficient to decrease both TEWL and total IgE. Aquaporin-3 gene expression, linked with skin hydration, was modulated in the skin of mice and human primary keratinocytes following protein hydrolysate exposure. Skin barrier improvement may be an additional mechanism by which pHF-W may potentially reduce the risk of AD development in infants. Further human studies are warranted to confirm the clinical efficacy of these observations.

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Patterned acquisition of skin barrier function during development

Development ◽

10.1242/dev.125.8.1541 ◽

1998 ◽

Vol 125 (8) ◽

pp. 1541-1552 ◽

Cited By ~ 16

Author(s):

M.J. Hardman ◽

P. Sisi ◽

D.N. Banbury ◽

C. Byrne

Keyword(s):

Stratum Corneum ◽

Barrier Function ◽

Skin Barrier ◽

Skin Barrier Function ◽

Skin Permeability ◽

Terrestrial Environment ◽

Additional Species ◽

Barrier Formation ◽

Whole Mount ◽

Moving Front

Skin barrier function is conferred by the outer layer of epidermis, the stratum corneum, and is essential for terrestrial life. Quantitative trans-epidermal water loss assays show that barrier forms late in embryogenesis, permitting the foetus to survive a terrestrial environment at birth. Using qualitative in situ assays for skin permeability, we show that barrier forms in a patterned manner late in mouse gestation. Barrier forms at specific epidermal sites, then spreads around the embryo as a moving front. The moving front of permeability change is accompanied by multiple changes in the outer, stratum corneum-precursor cells. We use the permeability assays to show that final stages of cornified envelope assembly are coordinated with initial stages of barrier formation. Hence the whole-mount permeability assays record developmental acquisition of a known, essential component of the adult barrier. We demonstrate the authenticity of the whole-mount assays after maternal glucocorticoid therapy (known to accelerate barrier formation) and in additional species including the rat where barrier formation is well characterized by TEWL assay (Aszterbaum, M., Menon, G. K., Feingold, K. R. and Williams, M. L. Pediatr. Res. 31, 308–317). The demonstration of patterned barrier formation in other species suggests patterned change as the universal mode of embryonic barrier acquisition. These results highlight the importance of patterning as a mode of epidermal maturation during development.

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The Role of Clindamycin Phosphate Associated with Adapalene in Three Semisolid Formulations Developed for Topical Acne Treatment

Revista de Chimie ◽

10.37358/rc.17.5.5585 ◽

2017 ◽

Vol 68 (5) ◽

pp. 937-943

Author(s):

Stela Mariana Al Hussein ◽

Nicoleta Todoran ◽

Silvia Imre ◽

Hussam Al Hussein ◽

Ana Melero Zaera ◽

...

Keyword(s):

Barrier Function ◽

Antimicrobial Agents ◽

Acne Vulgaris ◽

Skin Barrier ◽

Human Epidermis ◽

Skin Barrier Function ◽

Clindamycin Phosphate ◽

Oil In Water ◽

Study Results

Despite the fact that in mild-to moderate acne vulgaris the standard first-line therapy is the topical treatment with fixed combinations of antimicrobial agents and retinoids, the skin type and the skin barrier function should be taken into account when formulating a topical product. The aim of this study was the comparison of three new semisolid formulations developed for topical application by evaluation of their rheological behavior, as well as the evaluation of in vitro percutaneous diffusion through human epidermis membrane of the pharmaceutical ingredients. Clindamycin phosphate and adapalene were incorporated in three different topical bases, an HPLC method for the determination of their content in the new formulations being developed and validated. A higher concentration of drugs was released from the two gel systems (hydroxypropylmethylcellulose 2.5% -F1 and hydroxyethylcellulose 3% -F2) than from the oil-in-water cream (F3) at pH 7.4, whereas at pH 5.5 the drugs were released in higher amounts from the formulation F3. Following the rheological behavoir associated with the penetrability through the human epidermis membrane, our study results suggest that F1 and F2 could be appropriate in treating acne lesions in patients with oily skin and unaffected skin barrier function. In contrast, the oil-in-water cream (F3), due to its possible emolient effect and its higher penetrability at pH 5.5 than gel vehicles, may be indicated for patients with dry and sensitive skin associated with an altered skin barrier.

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